Recombinant Human beta-Nerve Growth Factor

 ( rHub-NGF)

Certificate of Analysis and Data Sheet

 

×                  Description:

Recombinant Human b-NGF produced in CHO is a homodimer, glycosylated, polypeptide chain containing 2 identical 119 amino acids and having a molecular mass of 16,950Dalton.

          The rHub-NGF is purified by proprietary chromatographic techniques.

 

×                  Source:

CHO cells.

 

×                  Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

 

×                  Formulation & Packaging:

The protein was lyophilized from a 0.2µm filtered solution in 30mM sodium acetate pH=5.5 containing 1% HSA.

 

×                  Solubility:

The lyophilized rHub-NGF is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=10.42).

 

×                  Stability:

Lyophilized rHub-NGF although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHb-NGF is best stored refrigerated at 4°C.

 

×                  Purity:

Greater than 99.0% as determined by:

 (a) Analysis by RP-HPLC.

           (b) Anion-exchange FPLC.

         (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained.

 

(Limit of acceptance:³98.0%. No more than 2% total impurities; no single impurity greater than 1%)

 

×                  Amino Acid Composition:

In total agreement with the expected amino acid composition of native human b-NGF .

 

×                  Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Ser-Ser-Ser-His-Pro, which agrees with the sequence of native human b-NGF.

 

×                  Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

 

×                  Biological Activity:

Our rHub-NGF is fully biologically active when compared to standards.

   The ED50,  calculated by PC12 cell line is 1.3-2 ng/ml and by DRG cell line is 0.8-1.5 ng/ml.

 

×                  Endotoxin:

Less than 0.1 ng/µg (IEU/µg) of rHub-NGF.

 

×                  Protein content:

         1.  Protein quantitation was carried out by two independent methods:

 

     UV spectroscopy at 280 nm.              

 

2.  Analysis by RP-HPLC, using a calibrated solution of b-NGF as a Reference Standard.

 

×                  Usage:

This material is offered for research, laboratory or further manufacturing purposes.